Transcellular transport of organic cations in double-transfected MDCK cells expressing human organic cation transporters hOCT1/hMATE1 and hOCT2/hMATE1
Transcellular transport of organic cations in double-transfected MDCK cells expressing human organic cation transporters hOCT1/hMATE1 and hOCT2/hMATE1
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DOI：
10.1016/j.bcp.2008.07.005
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发表时间：
2008-10-01
影响因子：
5.8
通讯作者：
Inui, Ken-ichi
中科院分区：
文献类型：
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作者：
Sato, Tomoko;Masuda, Satohiro;Inui, Ken-ichi
阐明有机阳离子通过基底外侧和顶端的跨细胞转运。为了研究人有机阳离子转运蛋白hOCT 1和hMATE 1的表达，我们建立了双转染的Madin-Darby犬肾（MDCK）细胞，分别表达人有机阳离子转运蛋白hOCT 1和hMATE 1（MDCK-hOCT 1/hMATE 1）以及hOCT 2和hMATE 1（MDCK-hOCT 2/hMATE 1）作为人肝细胞和肾上皮细胞的模型。使用特异性抗体，发现hOCT 1和hMATE 1或hOCT 2和hMATE 1分别定位于MDCK-hOCT 1/hMATE 1或MDCK-hOCT 2/hMATE 1细胞的基底膜和顶膜。在这些双转染子中，代表性底物[[C-14]四乙基铵从基底侧向顶侧单向转运。当顶侧的孵育介质的pH从5.5变化到8.5时，[C-14]四乙基铵的跨细胞运输的最佳pH被证明是6.5。在10 mM 1-甲基-4-苯基吡啶或1 mM左氧氟沙星存在下，两种双转染子的基底外侧的基底外侧至顶端的运输也减少。在MDCK-hOCT 2/hMATE 1细胞单层中，存在10 mM 1-甲基-4-苯基吡啶鎓时[14 C]四乙铵的蓄积减少，但存在1 mM左氧氟沙星时[14 C]四乙铵的蓄积显著增加，但在MDCK-hOCT 1/hMATE 1细胞单层中不存在。与对照细胞相比，HEK 293细胞对[C-14]四乙铵、[H-3]1-甲基-4-苯基吡啶、[C-14]二甲双胍和[H-3]西咪替丁的摄取受到hOCT 1、hOCT 2或hMATE 1表达的刺激，但对[C-14]普鲁卡因胺和[H-3]奎尼丁的摄取不受刺激。然而，[C-14]普鲁卡因酰胺和[H-3]奎尼丁的跨细胞转运在两个双转染子中清楚地观察到。这些细胞可用于检查化合物消除的途径，或预测转运蛋白介导的药物相互作用。(c)2008年爱思唯尔公司All rights reserved.
To clarify the transcellular transport of organic cations via basolateral and apical. transporters, we established double-transfected Madin-Darby canine kidney (MDCK) cells expressing both human organic cation transporter hOCT1 and hMATE1 (MDCK-hOCT1/hMATE1), and hOCT2 and hMATE1 (MDCK-hOCT2/hMATE1) as models of human hepatocytes and renal epithelial cells, respectively. Using the specific antibodies, hOCT1 and hMATE1 or hOCT2 and hMATE1 were found to be localized in the basolateral and apical membranes of MDCK-hOCT1/hMATE1 or MDCK-hOCT2/hMATE1 cells, respectively. A representative substrate, [[C-14]tetraethylammonium, was transported unidirectionally from the basolateral to apical side in these double transfectants. The optimal pH was showed to be 6.5 for the transcellular transport of [C-14]tetraethylammonium, when the pH of the incubation medium on the apical side was varied from 5.5 to 8.5. The basolateral-to-apical transport also decreased in the presence of 10 mM 1-methyl-4-phenylpyridinium or 1 mM levofloxacin on the basolateral side of both double transfectants. In MDCK-hOCT2/hMATE1 cell monolayers, but not in MDCK-hOCT1/hMATE1 cell monolayers, the accumulation of [14C] tetra ethylammonium was decreased in the presence of 10 mM 1-methyl-4-phenylpyridinium, but significantly increased in the presence of 1 mM levofloxacin. The uptake of [C-14]tetraethylammonium, [H-3]1-methyl-4-phenylpyridinium, [C-14]metformin and [H-3]cimetidine, but not of [C-14]procainamide and [H-3]quinidine, by HEK293 cells was stimulated by expression of the hOCT1, hOCT2 or hMATE1 compared to control cells. However, transcellular transport of [C-14]procainamide and [H-3]quinidine was clearly observed in both double-transfectants. These cells could be useful for examining the routes by which compounds are eliminated, or predicting transporter-mediated drug interaction. (c) 2008 Elsevier Inc. All rights reserved.